Thomas et al. 2025

Trifunctional lipid derivatives: PE’s mitochondrial interactome

Alix Thomas, Rainer Mueller, Ana Kojic, Scotland E. Farley, Frank Stein, Per Haberkant, Carsten Schultz

Correspondence: schulcar@ohsu.edu

Chemical Communications

https://doi.org/10.1039/D4CC03599B

Abstract

Phosphatidylethanolamine (PE) is a ubiquitous lipid species in higher eukaryotes which resides preferentially in mitochondria. Here, we synthesized a multifunctionalized PE derivative (1) de-signed to identify PE-binding proteins in intact cells through photo-crosslinking and subsequent isolation and proteomic analysis of the PE-protein conjugates. Due to its aromatic caging group, 1 is initially concentrating in perinuclear membranes. After uncaging, rapid translocation to mitochondria was observed. Hence, the tool is useful for tracking PE location and for determining the PE interactome. A trifunctional phosphatidic acid (PA) derivative in compari-son was rapidly metabolized and is hence more limited in its use.

Data visualization

Ranked-order Log2 Fold-change

Ranked-order plots depict the relative enrichment of each protein versus control (y-axis) from lowest to highest. The left panel depicts the results of a pulldown using trifunctionalized phosphatidic acid; the right panel depicts the results of the pulldown using trifunctionalized phosphatidylethanolamine. Black proteins are unenriched or depleted in the presence of probe, Purple proteins are enriched to the probe but fall below a FDR or p-value threshold of significance, and Orange proteins surpass the threshold of significance (need details of threshold).

Volcano plots

Volcano plots depict the relative enrichment of each protein versus control (x-axis) and the statistical significance of each protein following a Student’s t-test (y-axis, -log10 transformed). The left panel depicts the results of a pulldown using trifunctionalized phosphatidic acid; the right panel depicts the results of the pulldown using trifunctionalized phosphatidylethanolamine. Black proteins are unenriched or depleted in the presence of probe, Purple proteins are enriched to the probe but fall below a FDR or p-value threshold of significance, and Orange proteins surpass the threshold of significance (need details of threshold)

Enriched proteins

Data download